Flow cytometry is a well known technique that is used for high throughput measurements of optical and/or electrical characteristics of microscopic biological samples. Flow cytometry instruments analyse and isolate cells and organelles with particular physical, biochemical, and immunological properties.
Traditionally, cell sorting by flow cytometry (fluorescence activated cell sorting) has been the method of choice for isolation of specific cell populations by surface markers. However, cell sorting by flow cytometry suffers from several drawbacks, especially high dilution of desired cell sample, low speed and sterility problems. Furthermore, the equipment is very costly with high operation and maintenance cost, making the technique available only to a limited number of laboratories.
During the last few years, isolation of cells by antibody-coupled magnetic beads and carriers has been developed into a reliable tool for the isolation and characterisation of cell populations. Immunomagnetic cell separation, e.g. as commercially introduced by Dynal A/S and Miltenyi Biotec, has become an established method for cell analysis in clinical diagnostics. Due to the relatively low prize, this method is attractive in flow cytometry, especially in sorting of rare cellular events. For example, sorting of fetal cells contained in maternal blood sample provides a non-invasive alternative to prenatal diagnostic procedures, such as amniocentesis of chorionic villus sampling. Another rare event scenario is the detection of low concentration of cancer cells which has an important role in diagnosis of minimal residual disease and evaluation of appropriate therapies. Another medical application for cell sorting systems is the diagnosis of bacterial and viral diseases.
Although this method offers relatively inexpensive approach to sort rare cellular event, it adds considerable time onto the overall rare event detection and it does not offer the multiparameter analysis readily available with flow cytometry techniques. Existing techniques for magnetic separation are suffering from the low purity of the sorted cell fraction and the low recovery rate of the sorted cells. In most systems several washing steps have to be implemented into the separation procedure which then causes cell losses. Additionally small subpopulation of labelled cells cannot be directly isolated by existing magnetic separation techniques.
A good overview about fluorescence activated cell sorting procedures and magnetic activated cell sorting is given in Melamed et. al., “Flow Cytometry and Sorting, (Ed. Melamed et. al., Wiley & Sons Inc., 1990).